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RNase H

RNase H

Catalog Number: UA070045 Brand: UA BIOSCIENCE
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Regular price $84 USD
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Product Details

Product Specification


Synonyms Ribonuclease HI、RNase HI、Ribonuclease H、RNase H
Amino Acid Sequence

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Molecular Weight

19 kD

Purity >95% by SDS-PAGE
Conjugation Unconjugated
Tag His Tag
Physical Appearance Liquid
Storage Buffer

50 mM KCl、10 mM Tris-HCl、0.1 mM EDTA、1 mM DTT、200 µg/ml BSA 50% Glycerol(pH 7.4 @ 25°C)

Reconstitution

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Stability & Storage

Store at -25 ~ -15℃ for 2 years

Reference

 [1] Chan S H , Whipple J M , Dai N ,et al.RNase H-based analysis of synthetic mRNA 5' cap incorporation[J].RNA (New York, N.Y.), 2022, 28(8):1144-1155.

[2] Ilina T V , Brosenitsch T , Sluis-Cremer N ,et al.Retroviral RNase H: Structure, mechanism, and inhibition[J]. 2021. 

Background

Endonuclease that specifically degrades the RNA of RNA-DNA hybrids. RNase H participates in DNA replication; it helps to specify the origin of genomic replication by suppressing initiation at origins other than the oriC locus; along with the 5'-3' exonuclease of pol1, it removes RNA primers from the Okazaki fragments of lagging strand synthesis; and it defines the origin of replication for ColE1-type plasmids by specific cleavage of an RNA preprimer. Involved in production of retron derived msDNA (a branched RNA linked by a 2',5'-phosphodiester bond to a single-stranded DNA).

Components

Storage Solution: 5U/μl RNase H、50 mM KCl、10 mM Tris-HCl、0.1 mM EDTA、1 mM DTT、200 µg/ml BSA 50% Glycerol(pH 7.4 @ 25°C) 10*Reaction Buffer: 500 mM Tris-HCl、750 mM KCl、30 mM MgCl2、100mM DTT(pH 8.3 @ 25°C)

Protocol

Set-up a typical reaction as follows 1)Add the following components in sequence 2)Incubate at 37°C for 20 minutes This reaction can be scaled up according to experimental needs.

Guidelines

1、Both metal ion chelating agent and sulfhydryl sealer can inhibit RNase H activity 2、Heating at 65℃ can be inactivated for 10min

Unit Definition

One unit is the amount of enzyme required to hydrolyze 1 nmol nucleotide from a 20 pmol fluorescent-labeled 50 bp RNA-DNA hybridization chain at 37℃ for 20 minutes in a 50 µl reaction system.

Picture

Bioactivity

Marker(100 bp ~ 2000 bp):1 ul
Lane 1、DNA:RNA duplex Substrate
Lane 2、DNA:RNA duplex Substrate + Competitive product RNase H(5 U/ul)1 ul
Lane 3、DNA:RNA duplex Substrate + UA RNase H(5 U/ul)1 ul
Lane 4、DNA:RNA duplex Substrate + UA RNase H(5 U/ul)2 ul
Lane 5、DNA:RNA duplex Substrate + UA RNase H(5 U/ul)3 ul

SDS-PAGE

2μg (R: reducing condition, N: non-reducing condition).

SEC-HPLC

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